云南省德宏州居住区啮齿动物携带巴尔通体的调查研究

目的 调查云南省德宏傣族景颇族自治州（德宏州）居住区啮齿动物的巴尔通体感染和分布特点,为相关疾病防制提供参考依据。方法 采用笼夜法捕鼠,采集鼠类脾脏标本,用胰酶大豆琼脂培养基分离巴尔通体,用聚合酶链式反应法扩增巴尔通体gltA基因片段,并进行核苷酸序列测定和系统发育分析。结果 2011年在德宏州居住区共捕获啮齿动物5属9种340只和食虫动物2种18只,黄胸鼠为优势种（77.09%,276/358）。这些动物的脾脏标本中巴尔通体分离率为24.86%（89/358）,其中黄胸鼠分离率为28.99%（80/276）,其他阳性鼠种为黑缘齿鼠（5株）、社鼠（1株）、大绒鼠（1株）、小泡灰鼠（1株）和臭鼩鼱（1株）。共获得85株巴尔通体gltA基因核苷酸序列,进化分析表明,40株为特利波契巴尔通体（Bartonella tribocorum）,28株为伊丽莎白巴尔通体（B.elizabethae）,11株为昆州巴尔通体（B.queenslandensis）,3株为森林巴尔通体（B.silvatica）,3株未定种。黄胸鼠携带上述所有种类的巴尔通体。结论 德宏州啮齿动物中至少存在4种巴尔通体的流行,巴尔通体具有遗传多样性特点。以特利波契巴尔通体、伊丽莎白巴尔通体和昆州巴尔通体为主要流行菌株,黄胸鼠为当地居住区巴尔通体的主要宿主。今后应加强巴尔通体相关疾病的监测和防制工作。     

汉赛巴尔通体的分子分型研究进展

汉赛巴尔通体是一种新发的人兽共患病病原,呈全球性分布。随着分子生物学技术的发展,多种分子分型方法应用于其流行病学调查,加深了研究者们对汉赛巴尔通体种内变异和系统发育关系的了解。本文就几种常见的分子分型方法在汉赛巴尔通体分子流行病学研究中的应用做一综述。     

2017年云南省剑川县鼠形动物中巴尔通体感染状况调查

目的调查2017年云南省剑川县鼠形动物中巴尔通体的自然感染情况。方法2017年3—7月在剑川县用笼捕法捕获鼠形动物，无菌采集其股动脉血并提取DNA，应用荧光定量PCR技术进行巴尔通体检测，对检测阳性的血液样本进行巴尔通体分离培养。结果从剑川县沙溪镇石龙村及金华镇西门社区共捕获鼠形动物372只；经荧光定量PCR检测，阳性208份，阳性率为55.91%；11种鼠形动物中有10种检出阳性，其中大绒鼠的阳性率为38.73%，齐氏姬鼠为72.49%，贝氏树鼩为7.14%，中华姬鼠为77.78%，赤腹松鼠、社鼠、褐家鼠为50.00%，大足鼠、巢鼠为100.00%；阳性样本经分离培养后获得19株巴尔通体。结论TaqMan探针荧光PCR技术是巴尔通体感染的快速诊断方法。 剑川县鼠形动物中感染较高的为巴尔通体，受感染的鼠种较多，其中以齐氏姬鼠与大绒鼠较高，人群与外环境的鼠类接触存在感染发病的风险。     

滇西地区室内鼠形动物巴尔通体感染情况调查研究

目的 了解滇西地区室内鼠形动物巴尔通体感染状况。方法 在滇西地区8个州(市)抽取10个县(市、区)40个自然村,随机抽取800户家庭(每村20户),进行室内捕鼠,采集鼠脾脏标本并提取基因组DNA,运用聚合酶链反应(PCR)扩增巴尔通体枸橼酸合酶(gltA)基因,阳性者进行测序分析。结果 捕获9种421只鼠形动物,采集脏器样本404份,PCR扩增阳性64份,阳性率为15.84%。9种鼠形动物中仅黄胸鼠和褐家鼠检出阳性,阳性率分别为23.05%(62/269)和8.70%(2/23)。10个县(市、区)中6个检出阳性。基因分析表明携带的巴尔通体至少存在6种基因型:B. tribocorum、B. queenslandensis、B. elizabethae、B. rochalimae及2个可能的新型。结论 滇西地区室内鼠形动物感染巴尔通体,且呈现基因多样性特征,部分县(市)阳性率较高,需加强监测和预防控制。     

鼠形动物自然感染巴尔通体的研究进展

巴尔通体为革兰染色阴性杆菌，寄生于脊椎动物红细胞内，可引起巴尔通体病。随着交通、旅游业的发展和原生环境的开发，人类与鼠形动物的接触机会增多，可能导致巴尔通体病大范围传播，对人类健康造成潜在的威胁。本研究对巴尔通体的生物学特性、宿主动物、传播媒介以及流行现状等方面进行了综述。     

巴尔通体MALDI-TOF MS数据库的建立与鉴定方法的评价

目的建立巴尔通体的基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)数据库和质谱鉴定方法,利用野生菌株进行验证、评价。方法应用MALDI-TOF MS采集巴尔通体ATCC标准菌株和国内流行菌株的质谱数据,每株菌采集24张的质谱图集,获得特定的蛋白指纹图谱,汇总成不同种巴尔通体的标准质谱图,建立巴尔通体MALDI-TOF MS标准数据库。比较乙醇-甲酸提取法和直接涂抹法对质谱鉴定结果的影响,并进行重复性实验评估方法的稳定性。进一步应用173株野生巴尔通体菌株评估该标准数据库和相应的质谱鉴定方法。结果应用汉赛巴尔通体、五日热巴尔通体和杆菌样巴尔通体等21种/亚种巴尔通体,共计200株多地区来源的地理菌株,建立了巴尔通体MALDI-TOF MS蛋白指纹图谱标准数据库。2种样品制备方法均可以达到正确鉴定标准,提取法质谱得分略高于直接涂抹法。应用8种、16株巴尔通体菌株进行的重复性验证,得分为(9.078±0.031)~(9.776±0.006)分,变异系数(CV)为0.06%~1.12%。14种、173株巴尔通体测试菌株的质谱鉴定得分为9.014~9.796分,平均(9.462±0.195)分,在种水平上能被正确识别。结论基于该研究自主建立的巴尔通体质谱数据库,MALDI-TOF MS技术能够快速、准确鉴定巴尔通体种类,该课题组所建立的巴尔通体质谱标准数据库具有重要应用价值,对临床上巴尔通体病的实验室检验与诊断具有重要意义。     

新疆古尔图地区长尾黄鼠感染病原体情况调查

目的调查新疆维吾尔自治区（新疆）古尔图地区长尾黄鼠汉赛巴尔通体、伯氏疏螺旋体和嗜吞噬细胞无形体等病原体感染情况,分析该地区自然疫源性疾病流行风险,为制定防控措施提供科学依据。方法2017年5 — 9月采用一日弓形夹法捕捉查岗果勒、布兰布拉克、白石头等地长尾黄鼠86只,无菌采集鼠体肾脏,试剂盒法提取全基因组DNA;运用实时荧光定量聚合酶链式反应方法检测样本汉赛巴尔通体ssrA基因、伯氏疏螺旋体recA基因和嗜吞噬细胞无形体Msp2基因。结果共检测86只长尾黄鼠样本,其中汉赛巴尔通体阳性42份,阳性率为48.84%;伯氏疏螺旋体阳性6份,阳性率为6.98%;嗜吞噬细胞无形体阳性3份,阳性率为3.49%。 布兰布拉克地区的汉赛巴尔通体感染率高于查岗果勒和白石头地区,白石头地区的伯氏疏螺旋体和嗜吞噬无形体感染率高于其他两地。结论新疆古尔图地区长尾黄鼠存在汉赛巴尔通体、伯氏疏螺旋体和嗜吞噬细胞无形体感染。     

河南山东部分地区家猫汉赛巴尔通体感染情况调查

目的 调查河南、山东部分地区家猫汉赛巴尔通体血清抗体及菌血症情况.方法 间接酶联免疫吸附试验(ELISA)方法调查314份家猫血清抗体情况,通过聚合酶链反应(PCR)和测序对血培养产物进行病原学鉴定.结果 用ELISA法共检测314份标本,总的抗体阳性率为29.6%,＜6月龄的幼猫阳性率较低(19.3%),＞2岁的老猫抗体阳性率最高(37.5%),雌性和雄性之间没有明显差别,血培养共培养出42份可疑阳性,PCR及序列分析证实为汉赛巴尔通体.结论 河南、山东部分地区家猫均存在汉赛巴尔通体感染情况,汉赛巴尔通体是家猫感染的主要菌种.     

立克次体病的诊治方法进展

<正>立克次体是一类严格细胞内寄生的原核细胞型微生物,其中对人类治病的主要有5个属,包括立克次体科中的立克次体属、东方体属、柯克斯体属、埃里克体属以及巴尔通体科中的巴通体属。立克次体属又分为2个生物型:斑疹伤寒群和斑点热群。原有的恙虫病群已另列为东方体属,以及近年来逐渐多发的无形体属。立克次体感染性疾病在世界范围内分布广泛,目前我国流行的立克次体病主要有流行性斑疹伤寒、地方性斑疹伤     

猴株五日热巴尔通体药敏及生化特性研究

目的 了解猴株五日热巴尔通体的生化特性和药物敏感性, 为进一步研究五日热巴尔通体的耐药机制奠定基础。方法 采用E test法, 检测1株五日热ATCC参考株及10株五日热巴尔通体猴分离株的生化特性及对14种抗生素的最低抑菌浓度(MIC)。结果 H15SC对利福平高度耐药(MIC256), 其余菌株敏感;H98SC对克林霉素耐药, 其余菌株敏感;除S13外, 其余10株菌株对丁胺卡那霉素、多粘菌素MICs值较高;全部菌株对阿奇霉素、头孢他啶、环丙沙星、庆大霉素、红霉素、妥布霉素、氯霉素、强力霉素、苄星青霉素敏感。结论 首次发现巴尔通体菌株对利福平高度耐药, 需要进一步了解其耐药机制, 减少不合理用药, 预防利福平耐药在人分离株中发生。     

Culture-negative infectious endocarditis caused by Bartonella spp.: 2 case reports and a review of the literature

Bartonella spp. are rare pathogens in humans and were recently recognized as important causative agents of culture-negative endocarditis. Here, we describe the 1st 2 documented cases of culture-negative endocarditis due to Bartonella henselae and Bartonella quintana encountered in a single hospital in Germany. Infection of the heart valve tissue was detected by broad-range polymerase chain reaction (PCR) and further confirmed by serologic testing. In particular, acute B. henselae infection with an impressive bacterial colonization of the infected cardiac valve was illustrated by transmission electron microscopy. B. henselae was further characterized by PCR assays targeting genotype-specific regions. Disease progression was initially monitored over the entire infection episode through inflammatory markers. In addition, a short overview of published detailed cases of Bartonella endocarditis in Europe within the last 7 years is given.     

THE EFFECT OF BILATERAL SUPRARENALECTOMY IN ADULT ALBINO RATS ON THE NATURAL AND ACQUIRED RESISTANCE TO BARTONELLA MURIS ANEMIA

Normal adult Wistar rats (non-carrier stock) are readily infected with Bartonella muris and develop a severe anemia if large amounts of infecting material are used. The normal adult rat of Wistar stock possesses a relatively high natural resistance to spontaneous infection with this organism. Bilateral suprarenalectomy in Wistar rats lowers the natural resistance to a subsequent infection with Bartonella muris. This procedure does not alter the type of tissue response to the virus but lowers the natural resistance of the rat to toxic effects of the infection. The acquired immunity to Bartonella muris conferred by a first infection is not broken down by subsequent suprarenalectomy. The mechanisms of acquired and natural resistance are dependent on different physiological processes in the organism and are not merely quantitative variations of the same process as is generally assumed.     

Bartonella adhesin a mediates a proangiogenic host cell response

Bartonella henselae causes vasculoproliferative disorders in humans. We identified a nonfimbrial adhesin of B. henselae designated as Bartonella adhesin A (BadA). BadA is a 340-kD outer membrane protein encoded by the 9.3-kb badA gene. It has a modular structure and contains domains homologous to the Yersinia enterocolitica nonfimbrial adhesin (Yersinia adhesin A). Expression of BadA was restored in a BadA-deficient transposon mutant by complementation in trans. BadA mediates the binding of B. henselae to extracellular matrix proteins and to endothelial cells, possibly via beta1 integrins, but prevents phagocytosis. Expression of BadA is crucial for activation of hypoxia-inducible factor 1 in host cells by B. henselae and secretion of proangiogenic cytokines (e.g., vascular endothelial growth factor). BadA is immunodominant in B. henselae-infected patients and rodents, indicating that it is expressed during Bartonella infections. Our results suggest that BadA, the largest characterized bacterial protein thus far, is a major pathogenicity factor of B. henselae with a potential role in the induction of vasculoproliferative disorders.     

In vitro Bartonella quintana infection modulates the programmed cell death and inflammatory reaction of endothelial cells

Bartonella quintana is an epicellular bacterium, which in vivo as well as in vitro, invades endothelial cells and develops within them inducing proliferative effects that play a pivotal role in neovascular manifestation of this disease. We investigated the effect of live Bartonella quintana and its LPS on apoptosis and inflammatory response in HUVEC-C, an endothelial cell line. The kinetics of the programmed cell death of Bartonella quintana-infected HUVEC-C showed a peculiar course. Even if early during infection apoptosis reached a peak after 6 h, later on apoptosis was inhibited. Such apoptosis inhibition was not observed during Bartonella quintana lipopolysaccharide treatment because LPS-stimulated HUVEC-C did progress to cell death. Evaluation of multiple cell signal transduction pathways revealed an overexpression of Apaf 1 and caspase 8 in HUVEC-C after 2 h of infection, and of bcl-2 starting from 10 h post Bartonella quintana infection. Moreover, Bartonella quintana and its LPS showed a different effect on the activation of genes involved in inflammatory response as revealed by molecular analysis of host cells. Bartonella quintana appears to be able to inhibit programmed cell death, inducing intracellular signals leading to survival and proliferation through the bcl-2 gene, despite the early increase of inflammatory status induced in endothelial cells. This mechanism, together with a poor endotoxin ability to stimulate strong inflammatory response, could contribute to the capability of the bacteria to persist intracellularly, causing chronic disease and producing neovascular manifestations.     

Bartonella infection in hematological practice

AIM: To characterize the clinical and histological features of Bartonella infection in patients asking for hematological advice and to assess the significance of serological and molecular methods for the diagnosis of this infection. MATERIALS AND METHODS: The case histories of 747 patients asking for advice at the Hematology Research Cancer, Russian Academy of Medical Sciences, for lymphadenopaphy were retrospectively studied. The study included 10 patients in whom Bartonella infection could be suspected. For verification of the diagnosis, the authors conducted a serological study of the patients' sera and a molecular study of archival paraffined lymph node biopsy specimens. RESULTS: The study showed it possible to make a retrospective diagnosis of cat-scratch disease (CSD) by polymerase chain reaction (PCR) used in the study of archival lymph node biopsy specimens and stained preparations. CONCLUSION: CSD should be suspected when a patient has sustained lymphadenopathy and a respective epidemiological history (feline contact). Bartonella infection should be diagnosed on the basis of a dynamic serological study and, if possible, PCR of cells from biopsy specimens of lymph nodes or the lesion developed at the site of Bartonella penetration into the human body (primary affect).     

Subtyping of uncultured bartonellae using sequence comparison of 16 S/23 S rRNA intergenic spacer regions amplified directly from infected blood

This study aimed to assess the usefulness of a PCR-based approach to the detection and differentiation of Bartonella strains in infected blood. The conservation of potential genus-specific PCR primer hybridisation sites within the 16 S/23 S rRNA gene intragenic spacer regions of Bartonella species was confirmed following sequence analysis of the intragenic spacer regions of four previously untested species. The extent of intra-species variation within the specific amplicons was assessed by comparison of sequences obtained from 17 strains of four Bartonella species. Eight sequence variants were obtained. Each species for which multiple strains were tested possessed at least two intragenic spacer regions variants, but the differences between these strains were markedly less than those observed inter-species. Sequence analysis was performed on 60 amplicons obtained from blood pellets collected from woodland rodent communities in which bartonella infections were known to be highly prevalent. Twelve variants were encountered, only five of which had been found among the studied isolates. Partial intragenic spacer region amplification followed by product sequence analysis offers a potentially sensitive and totally transferable means of inter- and intra-species differentiation of Bartonella strains, and its use in this study has broadened our knowledge of the genotypic spectrum of bartonellae associated with natural infections among UK woodland rodents.